I am attempting to stimulate mouse T-cell proliferation in vitro using plate-bound CD3 (10ug/ml, clone 145-2C11) and soluble CD28 (2ug/ml, clone 37.51) antibodies. I can induce some activation as assessed by forward and side scatter parameters, but not to levels I would expect. It turns out that mice of the 129 background (for which my model is on) have an amino acid substitution within CD3e that compromises binding of the 145-2C11 clone. This leads to suboptimal activation. See reference https://www.ncbi.nlm.nih.gov/pubmed/20638133 for the data on this.
Does anyone know of another mouse-specific CD3e antibody that can be used to induce in vitro proliferation of T-cells or perhaps a way around this? Or, does anyone know if any of the human CD3e antibodies (OKT3, UCH1, etc) can induce proliferation of mouse cells? Thanks for any insights you can provide.
You can use Concanavalin A (Con-A) as an alternative for anti-CD3/CD28
There's the KT3 mAb clone, I've never used it but I've seen it used in literature. Miltenyi and Thermofisher both have bead-bound anti-mouse CD3/28 products. Not sure what anti-CD3 clones those use but that's another option.
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology
- Immunology Techniques